成纤维细胞生长因子1通过改善线粒体氧化应激缓解对乙酰氨基苯酚所致小鼠急性肝损伤  

Fibroblast growth factor 1 alleviates acetaminophen-induced acute liver injury in mice via improving mitochondrial oxidative stress

作　　者：彭紫颖 南燕[2] 谢先海[3] 李晓晓 高玲 张磊[1] 姚瑞娜 应磊 汪洋[1] PENG Ziying;NAN Yan;XIE Xianhai;LI Xiaoxiao;GAO Ling;ZHANG Lei;YAO Ruina;YING Lei;WANG Yang(Department of Pathophysiology,Wenzhou Medical University,Wenzhou 325035,China;Department of Neonatology,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China;Department of Trauma and Emergency,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China)

机构地区：[1]温州医科大学病理生理学教研室,浙江温州325035 [2]温州医科大学附属第二医院育英儿童医院新生儿科,浙江温州325027 [3]温州医科大学附属第一医院创伤急诊外科,浙江温州325015

出　　处：《温州医科大学学报》2023年第4期259-268,共10页Journal of Wenzhou Medical University

基　　金：浙江省自然科学基金项目(LQ22H020004);温州市基础性科研项目(Y2020228,Y20210694)。

摘　　要：目的:探讨成纤维细胞生长因子1(FGF1)缓解对乙酰氨基苯酚(APAP)所致小鼠急性肝损伤的机制。方法:①将C57BL/6J小鼠随机分为Control组、APAP 3 h组、APAP 6 h组、APAP 12 h组和APAP 24 h组,500 mg/kg剂量APAP腹腔注射造模后于对应时间点处死小鼠,肝脏DHE染色和F4/80染色观察活性氧(ROS)含量以及炎症细胞数目确定APAP小鼠急性肝损伤最显著时间点;②将C57BL/6J小鼠随机分为Control组、APAP组、APAP+FGF11 mg/kg组、APAP+FGF12 mg/kg组和APAP+FGF15 mg/kg组,500 mg/kg剂量APAP造模1 h后,腹腔注射对应剂量FGF1,于造模后6 h处死小鼠,通过肝脏HE染色观察损伤面积,酶标仪法检测谷丙转氨酶(ALT)、谷草转氨酶(AST)、超氧化物歧化酶(SOD)和丙二醛(MDA)含量,qPCR检测过氧化氢酶(CAT)、SOD2、核因子E2相关因子2(NRF2)、趋化因子配体5(CCL5)、肿瘤坏死因子α(TNFα)和白细胞介素-6(IL-6)基因表达,确定FGF1缓解APAP小鼠急性肝损伤的最优剂量;③将C57BL/6J小鼠随机分为Control组、3 h APAP组、3 h APAP+FGF1组、6 h APAP组和6 h A PAP+FGF1组,500 mg/kg剂量APAP造模1 h后,腹腔注射2 mg/kg剂量FGF1,于造模后对应时间点处死小鼠,酶标仪法检测ATP含量,Western Blot检测线粒体氧化磷酸化蛋白相对表达量;④使用不同浓度APAP(0、1、5、10、15、20、40和60 mmol/L)处理HepG2细胞24 h,采用细胞毒性实验(CCK-8)检测细胞活性;⑤在20 mmol/L APAP的MEM基础培养基中,加入不同浓度FGF1,使FGF1终浓度为0、0.5、1、5和10 mg/mL,处理24 h后CCK-8实验检测细胞活性。结果:①在APAP所致小鼠急性肝损伤中,ROS含量在造模后3 h明显升高,6 h时达高峰;炎症细胞数目在6 h明显升高且达到最高,12 h和24 h开始下降。②在APAP所致小鼠急性肝损伤中,FGF1蛋白的表达水平皆明显下降(均P<0.01)。③在不同剂量的FGF1治疗后,肝脏损伤面积缩小,血清ALT和AST水平均显著下调(均P<0.01);肝脏组织ROS含量显著减少,MDA含量下调,SObjective:To explore the function and mechanism of fibroblast growth factor 1(FGF1)on alleviating acute liver injury induced by acetaminophen(APAP)in mice.Methods:①C57BL/6J mice were randomly divided as the control group,APAP 3 h group,APAP 6 h group,APAP 12 h group,and APAP 24 h group.After intraperitoneal injection of 500 mg/kg APAP,the mice were killed at the corresponding time points.The most significant time point of acute liver injury in APAP mice was determined by observing the content of reactive oxygen species and the number of inflammatory cells using liver DHE staining and F4/80 staining.②C57BL/6J mice were randomly divided as the control group,APAP group,APAP+FGF11 mg/kg group,APAP+FGF12 mg/kg group,and APAP+FGF15 mg/kg group.After 1 h of modeling with a 500 mg/kg dose of APAP,the corresponding dose of FGF1 was intraperitoneally injected.The mice were euthanized 6 h after modeling.The damage area was observed by HE staining of the liver.Liver function level was detected by ALT and AST kits.Oxidative stress levels of the liver were detected by DHE straining,MDA and SOD kits,and gene expression of CAT,SOD2 and NRF2.Inflammation level of the liver was detected by F4/80 staining,and gene expression of CCL5,TNFαand IL-6.Based on the above results,the optimal FGF1 dose for the treatment of acute liver injury induced by APAP was determined;③C57BL/6J mice were randomly divided as the control group,3 h APAP group,3 h APAP+FGF1 group,6 h APAP group,6 h APAP+FGF1 group.After modeling with a 500 mg/kg dose of APAP for 1 h,a dose of 2 mg/kg FGF1 was administered intraperitoneally.The mice were euthanized at the corresponding time points after modeling.Then the reagent kit was used to detect ATP content,and Western blot was used to detect the relative expression level of Total Oxphos(CV-ATP5A,CIII-UQCRC2,CIV-MTCO1,CII-SDHB,and CII-NDUFB8)and 3NT proteins.Based on the above results,the mechanism of FGF1 alleviating acute liver injury induced by APAP was investigated;④HepG2 cells were treated with different

关 键 词：成纤维细胞生长因子1 对乙酰氨基苯酚 急性肝损伤 线粒体 氧化应激 

分 类 号：R915[医药卫生—微生物与生化药学]